Serdal ARSLAN, Ayşen AÇIKKOL, Ertan Mahir KORKMAZ

The optimization of aDNA extraction protocol and sex determination of Bronze Age individuals from Oylum Höyük (Kilis, Turkey)

Oylum Höyük falls within the borders of the province of Kilis, located in the southern part of Turkey. It is an important Bronze Age settlement dating back to 5500 BC. In this study, the sex of 36 individuals found in Oylum Höyük was examined using morphological and genetic methods. In the morphological method, sex was identified in 14 female and 13 male individuals. The remaining 8 individuals could not be identified due to the poor preservation of their skeletal remains. Genetic sex was determined with the amplification of sex-specific X, Y centromeric alphoid repeats and ZFX-ZYF gene regions of DNA, mostly extracted from teeth. Based on ZFX-ZYF gene regions, sex was successfully determined using the PCR-RFLP method. The amplification results of ZFX-ZYF gene regions were also tested by applying the PCR-RFLP method. The amplification of X, Y centromeric alphoid repeats was successful in 29 individuals, while sex determination based on ZFX-ZFY fragments was carried out in 22 individuals. The vast majority of the results determined by analysis of ancient DNA were consistent with the morphological method. However, these 2 methods produced slightly different results for 5 individuals. Finally, the results indicated that ancient DNA analysis is a reliable method for sex determination, especially in poorly preserved and fragmented skeletal remains.

Anahtar Kelimeler:

Key words: Ancient DNA, sex determination, Bronze Age, Oylum Höyük

The optimization of aDNA extraction protocol and sex determination of Bronze Age individuals from Oylum Höyük (Kilis, Turkey)

Keywords:

Key words: Ancient DNA, sex determination, Bronze Age, Oylum Höyük,

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Özgen E. Gaziantep-Kilis Regional research survey of the Höyük: Oylum Höyük Excavation Results Meeting. 4: 239-248, 1987. 2. Özgen E. Oylum Höyük, 1987. Excavation Results Meeting. 10: 95-102, 1989. 3. Özgen E. Oylum Höyük, 1988. Excavation Results Meeting. 11: 203-210, 1990.
Özgen E, Tekin H, Ensert HK. Oylum Höyük excavation studies in 1991 and 1993. Excavation Results Meeting. 16: 95- 105, 1995.
Özgen E, Helwing B, Engin E et al. Oylum Höyük, 2002. Excavation Results Meeting. 25: 339-344, 2004.
Özgen E, Engin A, Acara Eser M et al. Oylum 2006: Mozaikli Bazilika Excavation. Excavation Results Meeting. 29: 411-426, 2008. 7. Matheson CD, Loy TH. Genetic sex identifi cation of 9400-year- old human skull samples from Çayönü Tepesi, Turkey. J of Archaeo Science 28: 569-575, 2001.
Sivagami S, Rao AR, Varshney UA. Simple and cost-eff ective method for preparing DNA from hard tooth tissue, and its use in polymerase chain reaction amplifi cation of amelogenin gene segment for sex determination in an Indian population. Forensic Sci lnt 110: 107-115, 2000.
Eliášová I, Mazura I, Smejtek L. DNA analysis of ancient skeletal remains. Folia Biol (Praha) 56: 47–50, 2010.
Workshop of European Anthropologist. Recommendations for age and sex diagnoses of skeletons. J Hum Evol 9: 518-549, 1980.
Lovejoy CO, Meindl RS, Pryzbeck TR. Chronological metamorphosis of the auricular surface of the Ilium: A new method for the determination of adult skeletal age at death. Am J Phys Anthropol 68: 15-28, 1985.
Kaur H, Jit I. Age estimation from cortical index of the human clavicle in Northwest Indians. Am J Phys Anthropol 83: 297- 305, 1990.
Brown K. Ancient DNA applications in human osteoarchaeology: achievements, problems and potential. In: Cox M. and Mays S. (eds.) Human Osteology in Archaeology and Forensic Science. Greenwich Medical Media, London; 2000: pp. 455-473.
Bouwman AS, Chilvers ER, Brown KA et al. Brief communication: identifi cation of the authentic ancient DNA sequence in a human bone contaminated with modern DNA. Am J Phys Anthropol 131: 428-431, 2006.
Kemp BM, Smith DG. Use of bleach to eliminate contaminating DNA from the surface of bones and teeth. Forensic Sci Int 15: 453-61, 2005.
Sambrook J, Fritsch E, Maniatis T. Molecular Cloning - A Laboratory Manual. 2nd Ed. Cold Spring Harbor Laboratory Press. New York; 1989.
Kaestle FA, Horsburgh KA. Ancient DNA in anthropology: Methods, applications, and ethics. Am J Phys Anthropol 35: 92-130, 2002.
Lin Z, Kondo T, Minamino T et al. Sex determination by polymerase chain reaction on mummies discovered at Taklamakan desert in 1912. Forensic Sci Int 75: 197-205, 1995.
Stacks B, Witte M. Sex determination of dried blood stains using the polymerase chain reaction (PCR) with homologous X-Y primers of the zinc fi nger protein gene. J Forensic Sci 41: 287-290, 1996.
DeGusta D, White TD. Technical report on the use of skeletal collections for DNA analysis. Ancient Biomolecules 1: 89-92, 1996.
Alvarez GA, Munoz I, Pestoni C et al. Eff ect of environmental factors on PCR-DNA analyses from dental pulp. Int J Leg Med 109: 125-129, 1996.
Kemp BM, Monroe C, Smith DG. Repeat silica extraction: A simple technique for the removal of PCR inhibitors from DNA extracts. J Archaeol Sci 33: 1680-1689, 2006.
Scholz M, Giddings I, Pusch CM. A polymerase chain reaction inhibitor of ancient hard and soft tissue DNA extracts is determined as human collagen type I. Anal Biochem 259: 283- 6, 1998.